| Objective:This study investigated the effects of acupuncture motor zone on neurological recovery,cerebral infarct area,pathological morphological structure of brain tissue,apoptosis index,expression levels of BDNF/CREB pathway and apoptosis-related proteins(Bcl-2,Bax,Caspase-3)in brain tissue of MCAO model rats,and explored the effect of acupuncture motor zone on the treatment of ischemic stroke by regulating neuronal apoptosis.This study was conducted to investigate the mechanism of ischemic stroke treatment by acupuncture in the motor area through regulation of neuronal apoptosis,and to provide a basis and method for clinical prevention and treatment of ischemic stroke.Methods:Sixty SPF-grade SD rats were randomly divided into 20 in the sham-operated group,20 in the model group,and 20 in the needle-prick group.The sham-operated group was operated only by grasping anesthesia and isolating the right internal carotid artery,but no wire bolus was inserted.The ischemic stroke model was prepared in the model group and the acupuncture group by inserting a wire thrombus into the right internal carotid artery to permanently infarct the middle cerebral artery(MCAO).After the rats were awake,their neurological function was scored according to the Zea-Longa neurobehavioral scoring method.Models with scores of 1-3 were included in the experiment,and models with scores of 0 and 4 and rats that died before the treatment time were excluded.The acupuncture group was selected to treat rats with acupuncture in the motor area of the head.(The locomotor area was positioned with reference to the International Standardized Scheme of Scalp Acupuncture Point Names and the WISTAR Anatomical Atlas of the Rat for standard positioning.)After the locomotor area was marked by shaving and local routine disinfection,the locomotor area in the anterior temporal oblique line was needled with a 0.16×25 mm milli-needle,and the rats were allowed to move around with the needle after needling,and the needle was retained for 40 min,and the needle was performed once every 10 min,and the treatment was performed once a day for 7 days.The sham-operated group and the model group were only marked by grasping and shaving without treatment.After the last treatment,the rats in each group were scored according to the Zea-Longa scale for their neurological neurobehavior to compare the recovery of neurological function;after brain extraction,the area of cerebral infarction in each group was determined by TTC staining;the morphological structure of brain tissue in each group was observed by HE staining;the apoptotic index of neuronal cells was calculated by Tunel assay;Western Blot assay was performed in each group of rats The expression levels of BDNF/CREB pathway and apoptosis-related proteins(Caspase-3,Bax,Bcl-2)in the brain tissues of each group were measured by Western Blot.Results:(1)Zea-Longa neurobehavioral scores:Zea-Longa neurobehavioral score scores were elevated in rats in the model and acupuncture groups after modeling and in rats in the model group after treatment compared with the sham-operated group(P<0.01).The Zea-Longa neurobehavioral scores were decreased in the treated rats in the acupuncture group compared with the model group(P<0.01).(2)TTC staining:the infarct area of rats in the model group(37.75±52.52)mm~2,and the infarct area of rats in the needle group(24.00±24.49)mm~2,were higher than the infarct volume of rats in the sham-operated group(0.00±0.00)mm~2(P<0.01);the infarct volume of rats in the needle group was lower than that of the model group(P<0.05).(3)Hematoxylin-eosin(HE)staining:brain nerve cells in the sham-operated group were arranged in an orderly manner,the cell membrane was basically intact,and the nuclei were clear;nerve cells in the model group showed edema and disorder,and obvious nuclear consolidation and nucleolysis were seen;brain nerve cells in the acupuncture group were arranged in a relatively normal manner,and the disorder was less,the cell membrane was basically intact,and the nuclear consolidation and nucleolysis were significantly improved compared with the model group.The nucleus consolidation and nucleolysis were greatly improved compared with the model group.(4)Tunel assay results:the number of apoptotic cells in the model and acupuncture groups was more than that in the sham-operated group(P<0.01);the number of apoptotic cells in the acupuncture group was less than that in the model group(P<0.01).(5)Western Blot assay results:BDNF and CREB protein expression was significantly reduced in the model group compared with the sham-operated group,and the difference was statistically significant(P<0.01);BDNF protein expression was increased in the acupuncture group compared with the model group,and the difference was statistically significant(P<0.05).Compared with the model group,CREB protein expression was significantly increased in the acupuncture group,and the difference was statistically significant.Compared with the sham-operated group,Caspase-3 and Bax protein expression were significantly increased in the model group,and the difference was statistically significant(P<0.01);compared with the model group,Caspase-3 and Bax protein expression were significantly decreased in the acupuncture group,and the difference was statistically significant(P<0.01).Compared with the sham-operated group,Bcl-2 protein expression was significantly reduced in the model group,and the difference was statistically significant(P<0.01);compared with the model group,Bcl-2protein expression was increased in the acupuncture group,and the difference was statistically significant(P<0.05).Conclusions:Acupuncture in the motor zone can promote neurological recovery in rats with ischemic stroke;reduce the infarcted area of brain tissue;improve the morphological structure of brain tissue;and reduce apoptosis of neuronal cells in the ischemic hemispheric zone.The mechanism of action of acupuncture in the motor zone for the treatment of ischemic stroke model rats may be related to the inhibition of neuronal apoptosis by activating the BDNF/CREB signaling pathway,promoting Bcl-2protein,and inhibiting Caspase-3 and Bax protein expression. |
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